Project Summary/Abstract Alcohol use disorders (AUDs) affect millions of people and burdens society with large socioeconomic costs. Despite the socioeconomic and health impact of alcohol dependence, there are only three treatments currently approved for AUDs, which are only modestly effective. Currently, the molecular mechanisms contributing to neuroadaptive changes that result in alcohol tolerance, dependence, and withdrawal are not fully understood. Thus, understanding the molecular mechanisms underlying the transition from controlled drinking to alcohol dependence is crucial in order to identify novel drug targets to advance the treatment of AUDs. Recent evidence suggests that alcohol dysregulates pro-inflammatory toll-like receptor (TLR) signaling in the brain. Pharmacological or genetic manipulation of the TLR signaling pathway reduces alcohol intake and preference?suggesting that neuroimmune signaling could be an effective therapeutic target for the treatment of AUDs. Therefore, the proposed research will investigate the molecular mechanisms of an unexplored branch of the TLR signaling pathway (TRIF-dependent) and its role in alcohol abuse. The first aim will use mass spectrometry and phosphoproteomics to determine how alcohol alters neuroimmune kinase activity as well as the global kinome in brain. The second and third aim will use null mutants, viral-mediated gene manipulation and mouse behavioral models to test the functional role of TRIF-dependent signaling in regulation of ethanol consumption. Thus, the proposed studies were designed to investigate TRIF-dependent signaling as an untapped source of new targets for drug development and genetic study. This study will provide crucial new knowledge about an unexplored neuroimmune pathway and its role in alcohol addiction. Results from Aim 1 will determine global kinase dysregulation in alcohol use disorder that will facilitate development of new pharmacological targets. This study will also determine which signaling components of the TRIF-dependent neuroimmune signaling pathway are critical for regulation of ethanol consumption. By understanding how TRIF-dependent signaling can facilitate decreased alcohol consumption, results from this project will implicate interesting new pharmacological targets and facilitate the discovery of novel genes and pathways that might contribute to the initiation, maintenance, or progression of alcoholism. Furthermore, the results of this research may extend to other addictions and diseases that involve kinase dysregulation and neuroimmune signaling, such as ischemia, Alzheimer's Disease, Multiple sclerosis, Parkinson's disease, and traumatic brain injury. Thus, understanding common signaling pathways provides an opportunity for developing therapeutic agents that will work in the treatment of multiple conditions, thereby increasing the quality of life for individual patients.